BE1030385B1 - CAPSULES FOR ORAL USE COMPRISING A PROBIOTICS-BASED FORMULATION, PACKAGED AS A UNIT DOSE IN A LAMINATED BLUSTER PACKAGE - Google Patents

Abstract

The invention relates to capsules for oral use comprising a shell essentially made of HPMC and a formulation comprising i) at least 5 billion CFU coming from at least 5 probiotic strains, ii) at least one prebiotic, iii) at least one nutraceutical; said capsules being in unit dose in a blister pack having a thermoformed part forming several shells and consisting of a laminate of at least 3 successive PVC/PE/PVDC layers. The capsules of the invention provide benefits to the body, and make it possible to reduce the degradation of probiotics linked to (i) oxygen and humidity during their storage and (ii) stomach acidity. during their ingestion.

Description

; ‚ BE2022/5207

CAPSULES FOR ORAL USE COMPRISING A FORMULATION

BASED ON PROBIOTICS, PACKAGED IN A UNIT DOSE IN A

LAMINATED BLUSTER PACKAGING

The present invention relates to capsules for oral use containing probiotics, one or more prebiotics as well as one or more nutraceuticals. In particular, the invention relates to such capsules individually packaged in a blister pack having a thermoformed part forming several shells and consisting of a laminate of at least 3 layers. The capsules of the invention are intended to provide benefits to the body and have a long shelf life.

Probiotics, also called "good bacteria", are useful micro-organisms which constitute the oral, intestinal and vaginal flora. A probiotic bacteria is defined as a living micro-organism which, when administered in adequate quantities, can confer an advantage for the health of the host (FAO/WHO 2001). Their presence/ingestion makes it possible in particular to counter the proliferation of harmful micro-organisms which can, for example, cause infectious diarrhea or vaginitis. In A healthy organism, the digestive tract is colonized by approximately 100,000 billion bacteria belonging to 400 different species. However, certain factors can interfere with the work accomplished by these bacteria such as a change in diet, a weakening of the immune system, diarrhea, infection ( for example gastroenteritis), physical or emotional stress and taking antibiotics.

In these cases, probiotic supplements, depending on their strain, can promote the balance of oral, intestinal or vaginal flora, help increase natural defenses by fighting against the spread of bad bacteria, help treat and prevent diarrhea, improve the absorption of certain nutrients. Probiotic supplements for oral use often come in the form of capsules containing a formulation in a rigid or soft shell.

Two major challenges exist when probiotics are integrated into the formulation of a capsule for oral use: their lyophilization/hydration on the one hand, and resistance to stomach acidity on the other hand.

Indeed, firstly, the benefits of probiotics most often require that these organisms must be alive at the time they enter the intestinal tract. To be able to handle and store formulations containing probiotics for several months, it is necessary to dehydrate them. Water is a limit to the long-term stabilization of cells; it is what maintains the biochemical reactions necessary for life. Without water, the bacteria temporarily cease to live, their need for oxygen and sugars is interrupted, they are inactivated until they are rehydrated. Unfortunately, dehydrated probiotics are very sensitive to oxygen and humidity.

Once the probiotic cells are dehydrated, they find themselves in direct contact with the oxygen in the ambient air. The risk of oxidative stress is therefore at its maximum. Indeed, not only are dehydrated probiotic cells more exposed to reactive oxygen species, but they are also deprived of their antioxidant system.

The conservation of formulations including probiotics is directly linked to their dehydration factor. In a humid environment, dehydrated probiotics will tend to absorb water from the atmosphere in which they have been placed. The more humid the atmosphere, the more marked this tendency will be and the more the probiotics will become enriched in water and become fragile, making the formulation less effective. This problem arising from humidity in the atmosphere is all the more critical in hot climates with high humidity levels such as so-called “tropical” climates.

In particular, document EP3446695 describes soft gelatin capsules containing probiotics in which a balance is found between the water activity necessary for the revivification of the probiotics and the water activity harmful to the conservation of a formulation. of probiotics in capsules.

Secondly, when a conventional probiotic capsule is administered, it most often dissolves in the stomach due to the acidity of the environment. The probiotics are immediately rehydrated and therefore come out of their inactive state. Additionally, some strains of probiotics are also particularly sensitive to low pH and must be able to survive through stomach and bile acids to reach the gastrointestinal tract and thus provide their benefits.

Finally, it is also known to combine probiotics with prebiotics, also called foods with a prebiotic effect, which constitute food for intestinal bacteria: they do not need it to live, but prebiotics promote their growth and allow them to exercise their beneficial functions on health more effectively. In particular, document EP2315582 discloses a capsule containing a mixture of probiotics and prebiotics such as FOS. Each capsule according to this document includes 3 strains of probiotics (Bacillus subtilis, Bacillus coagulans and

Enterococcus faecium or faecalis) at a rate of 500 million CFU for each strain according to an example. The capsules may also contain yeast and/or vitamins such as vitamin C, B6, BI and

B2.

The invention aims to overcome the disadvantages of the state of the art by providing capsules for oral use containing a formulation based on probiotics providing benefits to the body, and making it possible to reduce the degradation of the linked probiotics. (i) to oxygen and humidity during their storage in a dehydrated state and (ii) to the acidity of the stomach during their ingestion.

An objective of the invention is to provide capsules for oral use containing a formulation based on probiotics providing benefits to the body, said capsules being packaged in unit doses in a blister pack. In particular, the benefits targeted by the invention are in particular an improvement in the functioning of the immune system OR a normalization/stabilization of sugar and insulin levels in the blood or a reduction in the risk of urinary infection or a normalization of the rate of cholesterol in the blood, or aiding the proper development of bones and the normal functioning of the immune system in children or maintaining normal functions of the neurological system.

Another objective of the present invention is to provide capsules for oral use containing a formulation based on probiotics, said capsules being packaged in unit doses in a blister pack, very resistant to the degradation of probiotics linked to oxygen. and humidity when stored in a dehydrated state, especially in hot and humid climates, thereby increasing their shelf life.

Another objective of the present invention is to provide capsules for oral use containing a formulation based on probiotics, said capsules being packaged in unit dose in a blister pack, making it possible to reduce the degradation of probiotics linked to acidity. of the stomach during their ingestion, thus maintaining their effectiveness in the intestine. 5 To achieve these objectives, according to the invention capsules for oral use are provided comprising an envelope essentially made of HPMC and a formulation; said formulation comprising: i) at least 5 billion CFU coming from at least 5 probiotic strains including Bacillus Coagulans in the form of spores, the other probiotic strains being chosen from Lactobacillus

Acidophilus, Lactobacillus Gasseri, Lactobacillus Plantarum,

Lactobacillus Reuteri, Bifidobacterium Breve, Bifidobacterium

Infantis, Bifidobacterium Longum, Lactobacillus Casei,

Lactobacillus Helveticus, Lactobacillus Paracasei, Lactobacillus

Salivarius, Lactobacillus Crispatus, Bifidobacterium Animalis

Lactis, Lactobacillus Fermentum, Lactobacillus Rhamnosus,

Saccharomyces boulardi, ii) at least one prebiotic chosen from inulin, FOS, 2'-fucosyllactose, 3-fucosyllactose and lactodifucotetraose, iii) at least one nutraceutical chosen from pine phytosterol, vitamin Bó, vitamin B9 , proanthocyanidins, nopal, chromium, vitamin C, vitamin D3 and zinc; said capsules being packaged in Unit dose in a blister pack having (a) a thermoformed part forming several shells and consisting of a laminate of at least 3 successive PVC/PE/PVDC layers, the PVC layer being the internal layer in contact with the capsules, each blister comprising a capsule and (b) an aluminum foil arranged to cover the rear of the thermoformed portion of the blister pack.

The invention is thus based on a new and inventive approach. In particular, the inventors found, surprisingly, that by combining capsules made of a particular envelope containing a specific formulation and a very particular blister packaging, it was possible to achieve the set objectives.

Other characteristics, details and advantages of the invention will emerge from the description given below, without limitation.

In the present description and the claims, it is understood that, as used herein, the terms "ONE", "one" or "the" mean "at least One" and should not be limited to "only ONE". , unless explicitly stated otherwise, Additionally, when a value range is indicated, the endpoints are included. Finally, all integral and subdomain values in the numeric range are expressly included as if explicitly written.

According to the invention, the capsules comprise an envelope essentially made of HPMC. By HPMC, as well known in the field, we mean hydroxypropylmethylcellulose, also commonly called vegetable hypromellose. HPMC has resistance to acidity which helps protect the formulation when the capsule passes through the stomach. Also advantageously, HPMC is a compound of plant origin, compatible with diets excluding all or certain product(s) of animal origin.

By envelope essentially made of HPMC, we mean an envelope made of HPMC which contains at least 90% by weight of HPMC. Preferably, the envelope of the invention comprises at least 93% by weight of HPMC, or even at least 95% by weight of HPMC. According to one embodiment, the envelope of the invention comprises titanium dioxide and/or gellan gum.

Most preferably, the envelope of the invention consists of HPMC only (with the exception of impurities or traces of other elements).

According to the invention, the formulation comprises at least 5 billion CFU from at least 5 probiotic strains including Bacillus

Coagulans in the form of spores, the other probiotic strains being chosen from Lactobacillus Acidophilus, Lactobacillus Gasseri,

Lactobacillus Plantarum, Lactobacillus Reuteri, Bifidolbacterium Breve,

Bifidobacterium Infantis, Bifidobacterium Longum, Lactobacillus Casei,

Lactobacillus Helveticus, Lactobacillus Paracasei, Lactobacillus Salivarius,

Lactobacillus Crispatus, Bifidobacterium Animalis Lactis, Lactobacillus

Fermentum, Lactobacillus Rhamnosus, Saccharomyces boulardii.

By CFU (or “CFU” OR “colony forming Unit” in English) we mean an estimate of the number of viable bacteria or fungal cells in a sample.

According to one embodiment, the formulation comprises at least 6 probiotic strains, preferably at least 7 probiotic strains, preferably at least 8 probiotic strains, preferably at least 9 probiotic strains, and most preferably at least 10 probiotic layers.

According to the invention, the formulation comprises at least one prebiotic chosen from inulin, FOS, 2'-fucosyllactose, 3-fucosyllactose and lactodifucotetraose. By FOS we mean fructo-oligosaccharides. Preferably, the prebiotic is inulin.

According to the invention, the formulation comprises at least one nutraceutical chosen from pine phytosterol, vitamin B6, vitamin B9, proanthocyanidins, nopal, chromium, vitamin €, vitamin D3 and zinc.

By nutraceutical, we commonly mean a biocompatible product or food having positive effects, real or claimed, on health.

By nopal, we mean cactus fruit extract or Opuntia ficus-indica.

By chromium is meant any biocompatible chromium salt such as chromium (III) chloride.

By zinc is meant any biocompatible zinc salt such as zinc bisglycinate.

According to the invention, the capsules are packaged in unit doses in a blister pack having a thermoformed part forming several shells and consisting of a laminate of at least 3 successive PVC/PE/PVDC layers, the PVC layer being the internal layer in contact with the capsules, each shell comprising a capsule.

By PVC we mean polyvinyl chloride. By PE we mean polyethylene. By PVDC we mean polyvinylidene chloride.

According to the invention, the thermoformed part of the blister packaging may comprise one or more additional layer(s), in addition to the 3 successive PVC/PE/PVDC layers. It can, for example, include one or more additional layer(s) between the PVC and the

PE, and/or between PE and PVDC, or even on the PVDC layer (on the exterior side of the laminate).

According to the invention, an aluminum sheet is arranged to cover the rear of the thermoformed part of the blister packaging and thus ensure the sealing of the shell comprising a capsule.

Advantageously, these capsules have a shelf life of at least 18 months under conditions of ambient temperature between 4 and 40°C and relative humidity of 60 to 75%. Preferably, they have a shelf life of at least 24 months under conditions of ambient temperature between 4 and 40°C and relative humidity of 60 to 75%.

The conservation studies which were carried out under these conditions of temperature and relative humidity have shown that the probiotics in said capsules in their packaging according to the invention are stable according to ICH standards (International Council for

Harmonization of Technical Requirements for Pharmaceuticals for Human

Worn). The present invention provides six advantageous embodiments, depending on the specific health benefits sought.

In a first particular embodiment (embodiment 1), the formulation comprises at least 9 billion CFU from the following 8 probiotic strains: Bacillus Coagulans in the form of spores, Bifidobacterium Breve, Lactobacillus Acidophilus,

Lactobacillus Casei, Lactobacillus Plantarum, Lactobacillus Reuteri,

Lactobaccilus Rhamnosus, Saccharomyces boulardii; and vitamin € and vitamin D3.

This particular embodiment helps the proper functioning of the immune system.

In a second particular embodiment (embodiment 2), the formulation comprises at least 9 billion CFU from the following 7 probiotic strains: Bacillus Coagulans in the form of spores, Biidobacterium Animal Lactis, Lactobacillus Acidophilus,

Lactobacillus Casei, Lactobacillus Gasseri, Lactobacillus Plantarum,

Lactobacillus Rhamnosus; and chrome and nopal.

This particular embodiment helps normalize and stabilize blood sugar and insulin levels.

In a third particular embodiment (embodiment 3), the formulation comprises at least 9 billion CFU from the following 7 probiotic strains: Bacillus Coagulans in the form of spores, Biidobacterium Animalis Lactis, Bifidobacterium

Longum, Lactobacillus Acidophilus, Lactobacillus Crispatus, Lactobacillus

Fermentum, Lactobacillus Rhamnosus; and proanthocyanidins.

This particular embodiment helps reduce the risk of urinary infection.

In a fourth particular embodiment (embodiment 4), the formulation comprises at least 9 billion CFU from the following 5 probiotic strains: Bacillus Coagulans in the form of spores, Lactobacillus Acidophilus, Lactobacillus Gasseri,

Lactobacillus Plantarum, Lactobacillus Reuteri; and pine phytosterol.

This particular embodiment helps to normalize cholesterol levels in the blood.

In a fifth particular embodiment (embodiment 5), the formulation comprises at least 4 billion CFU from the following 8 probiotic strains: Bacillus Coagulans in the form of spores, Bifidobacterium Infantis, Lactobacillus Acidophilus,

Lactobacillus Helveticus, Lactobacillus Reuteri, Lactobacillus Rhamnosus,

Saccharomyces boulardii; zinc and vitamin D3.

This particular embodiment helps in the proper development of bones and the normal functioning of the immune system in children.

In a sixth particular embodiment (embodiment 6), the formulation comprises at least 9 billion CFU from the following 10 probiotic strains: Bacillus Coagulans in the form of spores, Bifidolbacterium Breve, Bifidobacterium Infantis,

Bfidobacterium Longum, Lactobacillus Casei, Lactobacillus Casei,

Lactobacillus Helveticus, Lactobacillus Paracasei, Lactobacillus

Plantarum, Lactobacillus Rhamnosus, Lactobacillus Salivarius; and vitamin B6 and vitamin B9.

This particular embodiment helps maintain normal functions of the neurological system.

Preferably, in the blister pack, the PVC layer has a thickness between 60 and 280 µm, the PVDC layer has a thickness between 30 and 45 µm, and the PE layer has a thickness between 15 and 280 µm. 70um.

According to the invention, the envelope, essentially made of HPMC, consists of two independent parts, fitted one inside the other and capable of being unboxed.

Preferably, the shape of the envelope essentially in

HPMC is such that the capsule can be swallowed and/or opened and its contents spilled, for example on food. The possibility of opening the capsules and spilling their contents is particularly suitable for children or the elderly who may have swallowing problems.

The preferred dosage for the capsules of the invention for oral use is one to two capsules per day.

It is of course understood that the present invention is in no way limited to the embodiments described above and that modifications can be made without departing from the scope of the claims. It is further understood that the invention also encompasses all possible combinations of features and preferred features, described herein and cited in the claims.

Additionally, the following examples are provided for purposes of illustration and are not intended to limit the scope of this invention.

Examples

Resistance to acidity:

In Vitro studies have shown that the shell of the capsules according to the invention effectively resist the acidity of the stomach for one hour. This experiment was carried out using said capsules comprising a shell essentially made of HPMC and a formulation comprising a probiotic. These capsules were immersed for one hour in a solution comprising 2g/L of

NaCl, 3.2 g/L of Pepsin and 7/mL/L HCL (37%), allowing a pH of 1.2 to be reached and simulating stomach conditions. They were then immersed in a solution comprising 3g/L of bile acid, 0.9 g/L of pancreatin and a sufficient quantity of NaOH to obtain a pH of 6.88 and simulate intestinal conditions. The CFU measurements carried out on the capsules having undergone a one-hour bath in the solution simulating stomach conditions followed by a 1, 2 or 3 hour bath in the intestinal solution show that more than half of the initial probiotics are still alive.

Conservation :

To study the stability of the probiotics in said capsules in their packaging, conservation studies were carried out.

These follow the ICH zone IV standards (hot and humid/tropical zone). 4 storage conditions were studied and are listed in the following table:

PE emmerere eme

Climate 30°C + 2°C 60% RH+5% RH

The cold conditions were reproduced in a Laebherr GKv 6410 refrigerator.

The real, climatic and accelerated conditions were reproduced in a WEISS PHARMA 600 climatic chamber. To guarantee the reliability of the results, the study was carried out in parallel on 2 samples of each particular embodiment according to the invention (embodiments 1 to 6).

A count of the total flora is carried out at each interval indicated in the following table.

The total flora count at 30°C includes the number of colonies that grow per gram or per ml of product on plate count agar after 72 hours aerobic incubation at 30°C. For Petri dishes which are intended for counting, a series of dilutions is carried out. The culture medium is prepared so that most of the microorganisms present grow well at this temperature. The solid sample is dissolved in buffered peptone water. This counting method meets the EN ISO 4833-1 standard.

ICH standards tolerate a drop of 0.5 log(CFU) during a complete conservation study. No decline of more than 0.5 log(CFU) was observed for all samples after 18 months in the cold, real and climatic storage conditions and after 6 months in the accelerated conditions.

Claims (12)

1. Capsules for oral use comprising a shell essentially made of HPMC and a formulation; said formulation comprising |) at least 5 billion CFU coming from at least 5 probiotic strains including Bacillus Coagulans in the form of spores, the other probiotic strains being chosen from Lactobacillus Acidophilus, Lactobacillus Gasseri, Lactobacillus Plantarum, Lactobacillus Reuteri, Bifidobacterium Breve, Bifidobacterium Infantis, Bifidobacterium Longum, Lactobacillus Casei, Lactobacillus Helveticus, Lactobacillus Paracasei, Lactobacillus Salivarius, Lactobacillus Crispatus, Bifidobacterium Animalis Lactis, Lactobacillus Fermentum, Lactobacillus Rhamnosus, Saccharomyces boulardi, ij) at least one prebiotic chosen from inulin, FOS, 2 '- fucosyllactose, 3-fucosyllactose and lactodifucotetraose, iii) at least one nutraceutical chosen from pine phytosterol, vitamin B6, vitamin B9, proanthocyanidins, nopal, chromium, vitamin C, vitamin D3 and zinc; said capsules being packaged in unit doses in a blister pack having (a) a thermoformed part forming several shells and consisting of a laminate of at least 3 successive PVC/PE/PVDC layers, the PVC layer being the internal layer in contact with the capsules, each blister comprising a capsule and (b) an aluminum foil arranged to cover the rear of the thermoformed portion of the blister pack. 2. Capsules according to claim 1, characterized in that they have a shelf life of at least 18 months under conditions of ambient temperature between 4 and 40°C and relative humidity of 60 to 75%. 3. Capsules according to claim 2, characterized in that they have a shelf life of at least 24 months under conditions of ambient temperature between 4 and 40°C and relative humidity of 60 to 75%. 4. Capsules according to one of the preceding claims, characterized in that the formulation contains inulin as a prebiotic. 5. Capsules according to one of the preceding claims, characterized in that the formulation comprises at least 9 billion CFU from the following 8 probiotic strains: Bacillus Coagulans in the form of spores, Bifidobacterium Breve, Lactobacillus Acidophilus, Lactobacillus Casei, Lactobacillus Plantarum, Lactobacillus Reuteri, Lactobaccilus Rhamnosus, Saccharomyces boulardii; and vitamin € and vitamin D3. 6. Capsules according to one of claims 1 to 4, characterized in that the formulation comprises at least 9 billion CFU from the following 7 probiotic strains: Bacillus Coagulans in the form of spores, Bifidobacterium Animal Lactis, Lactobacillus Acidophilus, Lactobacillus Casei, Lactobacillus Gasseri, Lactobacillus Plantarum, Lactobacillus Rhamnosus; and chrome and nopal. 7. Capsules according to one of claims 1 to 4, characterized in that the formulation comprises at least 9 billion CFU from the following 7 probiotic strains: Bacillus Coagulans in the form of spores, Biidobacterium Animalis Lactis, Bifidobacterium Longum, Lactobacillus Acidophilus, Lactobacillus Crispatus, Lactobacillus Fermentum, Lactobacillus Rhamnosus; and proanthocyanidins. 8. Capsules according to one of claims 1 to 4, characterized in that the formulation comprises at least 9 billion CFU from the following 5 probiotic strains: Bacillus Coagulans in the form of spores, Lactobacillus Acidophilus, Lactobacillus Gasseri, Lactobacillus Plantarum, Lactobacillus Reuteri; and pine phytosterol. 9. Capsules according to one of claims 1 to 4, characterized in that the formulation comprises at least 4 billion CFU from the following 8 probiotic strains: Bacillus Coagulans in the form of spores, Bifidobacterium Infantis, Lactobacillus Acidophilus, Lactobacillus Helveticus, Lactobacillus Reuteri, Lactobacillus Rnamnosus, Saccharomyces boulardii; zinc and vitamin D3. 10. Capsules according to one of claims 1 to 4, characterized in that the formulation comprises at least 9 billion CFU from the following 10 probiotic strains: Bacillus Coagulans in the form of spores, Biidobacterium Breve, Bifidobacterium Infantis, Bfidobacterium Longum, Lactobacillus Casei, Lactobacillus Casei, Lactobacillus Helveticus, Lactobacillus Paracasei, Lactobacillus Plantarum, Lactobacillus Rhamnosus, Lactobacillus Salivarius; and vitamin B6 and vitamin B9. 11. Capsules according to one of the preceding claims, characterized in that, in the blister pack, the PVC layer has a thickness of between 60 and 280 um, the PVDC layer has a thickness of between 30 and 45 um , and the PE layer has a thickness of between 15 and 70 μm. 12. Capsules according to one of the preceding claims, characterized in that the envelope essentially made of HPMC consists of two independent parts, fitted one inside the other and capable of being unboxed.